Elaboration of methods for detection of Xanthomonas axonopodis pv. phaseoli on bean seeds

Abstract

Xanthomonas axonopodis pv. phaseoli detection on artificially inoculated bean seeds was investigated. The method of the International Seed Federation - ISF (2006) was used. It included bacteria extraction from seeds and isolation on semiselective media with pathogenicity test of the investigated isolates. ELISA and PCR were used for verification of results. The results showed that the semiselective media MT (Milk Tween Agar) and XCP1 (Xanthomonas campestris pv. phaseoli Agar) were very suitable for isolation of X. a. pv. phaseoli. Pathogenicity was confirmed on young bean plants. ELISA test and PCR confirmed that all investigated isolates and reisolates belong to the bacterium X.a.pv. phaseoli.

U radu je ispitivana mogućnost utvrđivanja prisustva bakterije Xanthomonas axonopodis pv. phaseoli na veštački inokulisanom semenu pasulja. Ispitivanja su vršena po metodi International Seed Federation (ISF), ekstrakcijom bakterija iz semena i izolacijom na poluselektivne podloge uz proveru patogenosti dobijenih izolata. Za potvrdu dobijenih rezultata korišćen je ELISA test i PCR. Prema dobijenim rezultatima ova bakterija se može uspešno izolovati na poluselektivne podloge MT (Milk Tween Agar) i XCP1 (Xanthomonas campestris pv. phaseoli Agar). Patogenost je dokazana na mladim biljčicama pasulja. ELISA testovi i PCR su potvrdili da svi ispitivani izolati i reizolati pripadaju bakteriji X.a.pv. phaseoli.